Active beta-globin gene transcription occurs in methylated, DNase I-resistant chromatin of nonerythroid chicken cells.
We report active, inappropriate transcription of the chicken beta A-globin gene in normal fibroblasts, cultured MSB cells, and brain. We were unable to detect ovalbumin gene transcription in these same tissues. Most of the globin gene transcripts were found to be truncated near the beginning of the gene, suggesting the existence of a premature termination process that is preferentially active under conditions of inappropriate transcription. The inappropriately transcribed beta A-globin gene chromatin remained DNase I resistant and highly methylated. Thus, the DNase I-sensitive conformation of erythrocyte beta A chromatin was correlated not with beta A transcription per se but with beta A expression. Although both transcribed and nontranscribed genes within the globin domain exhibited the same DNase I sensitivity in erythrocyte nuclei, a housekeeping gene active in erythrocytes exhibited a different level of DNase I sensitivity. However, this gene exhibited the same level of DNase I sensitivity in both erythrocytes and a cultured cell line. These observations are consistent with the proposal (G. Blobel, Proc. Natl. Acad. Sci. USA 82:8527-8529, 1985) that the DNase I sensitivity of a gene may reflect properties of chromatin related to cotranscriptional and posttranscriptional aspects of mRNA production rather than to transcription per se.
Lois
R
R
Department of Chemistry and Biochemistry, University of California, Los Angeles 90024-1569.
Freeman
L
L
Villeponteau
B
B
Martinson
H G
HG
D000818:Animals; D015139:Blotting, Southern; D001921:Brain / Q000502:physiology; D002467:Cell Nucleus / Q000502:physiology; D002478:Cells, Cultured; D002645:Chickens; D002843:Chromatin / Q000502:physiology* / Q000648:ultrastructure; D003850:Deoxyribonuclease I / Q000494:pharmacology; D005347:Fibroblasts / Q000502:physiology; D005786:Gene Expression Regulation; D005914:Globins / Q000235:genetics*; D066298:In Vitro Techniques; D008745:Methylation; D012333:RNA, Messenger / Q000235:genetics; D015183:Restriction Mapping; D014158:Transcription, Genetic
D002843:Chromatin; D012333:RNA, Messenger; D005914:Globins; D003850:Deoxyribonuclease I